Development of a multivalent enterovirus subunit vaccine based on immunoinformatic design principles for the prevention of HFMD.

Hand, foot and mouth disease (HFMD) is mainly caused by EV-A71 and CV-A16. An increasing number of cases have been found to be caused by CV-A10, CV-A6, CV-B3 and the outbreaks are becoming increasingly more complex, often accompanied by the prevalence of a variety of enteroviruses. Based on the principle of synthetic peptide vaccines, we applied immune-informatics to design a highly efficient and safe multivalent epitope-based vaccine against EV-A71, CV-A16, CV-A10, CV-A6 and CV-B3. By screening B-cells, HTL and CTL cell antigen epitopes with high conservativity and immunogenicity that have no toxic effect on the host, further analysis confirmed that the vaccine built was IFN-γ inductive and IL-4 non-inductive HTL cell epitopes and had population coverage corresponding to MHC molecular alleles associated with T-cell phenotype. The multivalent enterovirus vaccine was constructed to connect the 50 s ribosomal protein L7/L12 adjuvant and candidate epitopes sequentially through appropriate linkers. Then, the antigenic, allergen and physical properties of the vaccine were evaluated, followed by a secondary structure analysis and tertiary structure modeling, disulfide engineering, refinement and validation. Moreover, the conformational B cell epitope of the vaccine was analyzed. The stability of the TLR4/MD2/Vaccine complex and details at atomic level were investigated by docking and molecular dynamics simulation. Finally, in silico immune simulation and in vivo immune experiments were done. This study provides a high cost-effective method of designing a multivalent enterovirus vaccine protect against a wide range of enterovirus pathogens.